Classical Complement Pathway ELISA Kit [Human] (CAT#: CB-P059-K) Datasheet

Kit

The complement system plays an essential role in chronic, autoimmune and infectious disease. There are three pathways of complement activation, namely the classical, the alternative and the lectin pathway. Impaired complement activity causes humans to become susceptible to repetitive fulminant or severe infections and may contribute to development of autoimmune disease. Inappropriate activation of complement contributes to chronic inflammation and tissue injury. In vitro activation of the complement sequence leads to the consumption of complement components which, in turn, leads to a decrease in their concentration. Thus, the determination of complement proteins or complement activity is used to indicate whether the complement system has been activated by an immunologic and/or pathogenic mechanism. Both functional and immunochemical complement measurements are used to evaluate subjects when a complement-activating disease is suspected or an inherited deficiency is possible. The level of complement activity evaluated by functional assays such as Complement kit takes into account the rate of synthesis, degradation, and consumption of the components and provides a measure of the integrity of the pathways as opposed to immunochemical methods which specifically measure the concentration of various complement components.

This kit is an enzyme immunoassay (ELISA) used to determine the functional classic complement pathway in human serum. For research use only, not for diagnostic procedures.

The Complement Classical pathway Kit combines principles of the hemolytic assay for complement activation with the use of labelled antibodies specific for neoantigen produced as a result of complement activation. The amount of neoantigen generated is proportional to the functional activity of complement pathways. In the Complement CP kit, the wells of the microtitre strips are coated with specific activators of the classical pathway. This in combination with sample dilution buffer composition and serum dilution level ensure that only the Classical pathway is activated. During the incubation of the diluted serum in the wells, complement is activated by the specific coating. The wells are then washed and the amount of C5b-9 complex formed on the plate surface is detected Alternative pathway (C3, factor D, B, P) C3 C3b C3a Lectin pathway (MBL, MASP -2, C4, C2) Classical pathway (C1q, C1r, C1s, C4, C2) C4b2a C4b2a C3bBbP Terminal pathway (C5, C6, C7, C8, C9 C5b-C9 (MAC) COMPL CP310 RUO, LABEL-DOC-0030, 3.0 3 with a specific alkaline phosphatase labelled antibody to the C5b-9 neoantigen formed during MAC (Membrane Attack Complex) formation. After a further washing step, detection of specific antibodies is obtained by incubation with alkaline phosphatase substrate solution. The amount of complement activation correlates with the colour intensity and is measured in terms of absorbance (optical density (OD).